Notch signaling in human iPS-derived neuronal progenitor lines from Focal Cortical Dysplasia patients.

Focal Cortical Dysplasia (FCD), a common type of Malformations of cortical development, may result from an early disturbance in the migration and final arrangement of the cortical architecture of immature neurons. FCD type II is now known to be due to a post-zygotic somatic mutation that involves the mTOR and AKT pathways. The aim of the present study was to investigate the possible differences in neurogenesis and neurodifferentiation of iPSCs (induced pluripotent stem cells) from fibroblasts of individuals affected by FCD type II (2) and normal individuals (2). iPSCs were generated from skin fibroblasts of FCD individuals and healthy individuals. The reprogramming was done through the fibroblasts exposure to viral vectors containing the OCT4, KLF4, SOX2, and c-MYC genes and the clones were characterized by immunohistochemistry. iPSCs were neurodifferentiated and analyzed at the 14th, 22nd and 35th days. We also analyzed the cerebral cortex tissue, fibroblasts and iPSCs cells from the individuals. Through qRT-PCR, the expression of 4 genes involved in Notch signaling process were quantified. In general, individuals with dysplasia presented increase and decrease in the relative quantification in the most genes analyzed compared to control individuals in all processes and study groups. We suggest that, during embryonic neurogenesis, the neural precursor cells of FCD type II individuals present increase and decrease in gene expression in the Notch signaling pathway causing cortical formation disorders and can be seen as a candidate for the developmental changes observed in the cerebral cortex of individuals with FCD type II. This altered gene expression may be related to brain formation with dysplasia.

GC/MS analysis and potential cytotoxic activity of Calyptranthes grandifolia (O. Berg), Calyptranthes tricona (D. Legrand) and Myrciaria plinioides (D. Legrand) essential oil in RAW264.7 and CHO-K1 cells.

The search for new bioactive substances derived from natural products is daily growing. Among biologically active products used in therapeutic approaches, essential oils are described with wide range of therapeutic and pharmacological potential. Plants from Myrtaceae family have the presence of essential oils in its composition. Calyptranthes grandifolia, Calyptranthes tricona and Myrciaria plinioides species belong to this family and are used by the local population. However, there are no reports in the literature describing relevant characteristics about the potential and possible activities of these species. The aim of this study was to identify the main compounds, evaluate the antioxidant potential and investigate, in RAW264.7 and CHO-K1 cells, the cytotoxic activity of the essential oils from the leaves of C. grandifolia, C. tricona and M. plinioides plants, in order to ensure their use. The compounds were identified by GC-MS, antioxidant activity was determined by DPPH method and cytotoxicity was assessed by the Alamar Blue method, at 48 and 72h. The main compounds found in the essential oils were sesquiterpenes. None of the essential oils have antioxidant potential. In cytotoxicity assays, the essential oils from the plants in analysis showed moderate activity in the proposed conditions. The alterations observed between the data provided by the essential oils in question, between different cell lines, may be associated with their composition, suggesting action of minor compounds. These results may suggest that the essential oils from C. grandifolia, C. tricona and M. plinioides have considerable potential to be explored. Future studies will be conducted to obtain more information about the action pathway and potential of the identified compounds.